TW201215876A - System and method for detecting amyloid proteins - Google Patents

Abstract

In accordance with an embodiment of the invention, there is provided a device and method for detecting an amyloid protein in an eye of a mammal. A method comprises illuminating the eye with a light source having at least one of a wavelength property, a polarization property or a combination thereof, each appropriate to produce fluorescence in at least an amyloid-binding compound when the amyloid-binding compound is bound to the amyloid protein, the amyloid-binding compound having been introduced to the eye and specifically binding to the amyloid protein indicative of the amyloidogenic disorder; and determining a time decay rate of fluorescence for at least the fluorescence produced by the amyloid-binding compound bound to the amyloid protein, the determining permitting distinguishing of the presence of the amyloid-binding compound bound to the amyloid protein in the eye based on at least the time decay rate.

Description

201215876 VI. Description of the Invention: [Technical Field of the Invention] The present invention relates to an apparatus and method for detecting a mammalian eye. μ poor white [Prior Art] ~ - Straight need to detect early progression (four) disease. Early detection enabled 2 to be treated early, and it has been proven that early treatment yields a higher success rate in the treatment of multiple diseases. It has been found that the analysis of the human eye, in particular the lens of the eye, provides an indication of various types of diseases. For example, researchers have found steroidal amyloid peptides and aggregates on the nucleus of the lens of the eye of Alzheimer's disease (Alzhei, s disease, [AD]). See U.S. Patent No. 7,297,326 to G. Id Stein et al. Since the nucleus is only part of the thickness of the ^mm, the measurement obtained from this region of the lens requires precise position, detailed information, and fast access. This measurement is especially needed because the human eye is almost continuously moving even when the patient is gazing at the target being illuminated. It has been shown that the presence of beta-type amyloid peptides and their aggregates in the test mammalian eye and/or in the cortical lens region or an increase in the normal control value indicates that the test mammal has neurodegenerative disorders such as Temple powder-like illness) or at risk of developing the disease. There is an urgent need for systems and methods for allowing early detection of amyloidogenic disorders. [Invention] According to one embodiment of the present invention, there is provided a method for detecting amyloids in the eyes of mammals 201215876, such as starches comprising aggregates Protein method. In some embodiments, the detection of amyloid is an indication of a halllet-like condition. The method comprises illuminating the eye with a light source having at least one of wavelength characteristics, polarization characteristics, or a combination thereof, each of the properties being suitable for producing in the starch-binding compound when at least one starch-like binding compound binds to the amyloid-like compound Fluorescent' such starch-binding compounds have been incorporated into the eye and specifically bind to an amyloid indicative of an amyloidogenic disorder; receive light comprising fluorescence produced by illuminating the eye; and assay at least by binding to amyloid Time decay rate of fluorescent fluorescence produced by a starch-binding compound such that the assay allows discrimination in the presence of a compound such as amyloid-like compound in the eye based on at least the temporal decay rate. In other related embodiments, the method can further comprise determining the fluorescence intensity of the fluorescent light produced by at least the starch-binding compound bound to the amyloid-like protein. The amount of the starch-binding compound bound to the amyloid-like protein can be determined based on at least one of the intensity and the time decay rate. The method can further comprise measuring the position of the ocular interface (such as the lens capsule of the eye) based on the fluorescent signal caused by the natural glory emitted by the tissue of the eye. At least one region of the eye may be sampled using illumination of the light source. 'The sampling includes performing a measurement of the entire region or illuminating the different locations within the sampling region with the light source. The sampling different locations includes illuminating at least one point, plane, and Or volume a sampling may include sampling a different position of the sigma on more than one area of the eye, and performing a planar scan of the eye using the light source in a continuous plane along the depth & % (9) ϋ $. The position on the nucleus of the fixed eye can be based on: (i) the distance from a particular anatomical structure (such as the eye crystal 201215876 corpuscular interface or the corneal interface) or (ii) the change in the measured intensity measure (slope). Identifying the presence of a starch-binding compound that binds to an amyloid-like protein can include spontaneous fluorescence of a starch-binding compound that binds to a powder-like protein such as a powder and a background, and spontaneous fluorescence of other non-specific dry and unbound developers. The method may comprise identifying at least one of the presence and the other of the following: a starch-like binding compound; a starch-binding compound that binds to an amyloid-like protein; and an amyloid-like protein. The amyloid-like protein may comprise aggregates or pre-amyloid aggregates (including peptide Api 42 and/or Ah one-mer, dimer or higher oligomers). For example, the amyloid-like protein may comprise a beta-based starch. The amyloidogenic disorder may comprise Alzheimer's disease. In other related embodiments, the starch-like binding compound may comprise a molecular rotor, a Chrysamine and/or a Goldenamine derivative, a Congo red and/or a Congo red derivative starch-binding compound; G or a gold amine G derivative type starch-binding compound; a thioflavin tau or a thioflavin tau derivative type starch-binding compound; and a thioflavin 8 or a thioflavin S derivative type powder-binding compound. The method y comprises identifying the presence of a class of powders based at least on the Rauma assay. The method can further comprise determining an average number of photons having a specific decay rate in a particular eye region. 2. The rate of transfer of the powder-like compound to the eye towel, the spatial distribution of the powder-like compound delivered to the eye, and/or the concentration gradient of the starch-binding compound at the corneal interface of the eye, based on the fluorescing assay. In addition, the spatial distribution of the powder-like compound and/or the time distribution of the starch-like compound in the aqueous humor can be determined based on the detected fluorescence. The method can further comprise determining at least one dimension of the anatomical structure or substructure based on natural fluorescent excitation of at least a portion of an anatomical structure of the eye or substructure 6 201215876. Determining the at least one dimension can comprise at least one of determining the thickness of the structure or substructure, determining the shape of the structure or substructure, and determining the distance between one or more structures or substructures of the eye. For example, determining the at least one dimension can include determining a corner thickness, a corneal shape, an aqueous humor depth, a lens shape, or a lens thickness' or determining an internal measure within the lens or other structure or substructure of the eye, such as a lens surface and The distance from the cortex or the nucleus or nucleus. The #法可进一# includes detecting the fluorescence generated by the eye using a photodetector device such as a photodiode, a photomultiplier, a charge coupled device (CCD), and an enhanced charge blessing device (ICCD). At least one of; for example, a rapid spur photodiode detector. The method can include performing a time-dependent single photon count of fluorescence produced by the eye. The time-dependent single photon count can include pulsing the source and over time The photon count distribution of the channel unit variation determines the time decay rate of the camp light. In other related embodiments, the induced natural fluorescence is measured and at least one of the regions of interest in the eye is determined from the method that the scan can be included in the eye; Detecting at least one region of interest of the sampling eye using a light source, the sampling comprising performing at least one complete region of at least one region or measuring at least one of different locations within the region using at least one of the regions - sampling different locations comprising Irradiating at least one of a point, a plane, or a volume in at least one region; The fluorescence intensity in the region combined with the fluorescence of the fluorescing and the sample in the fluorescein are used to determine the decay rate of light produced by at least a starch-binding compound such as at least a protein. For example, the method of 201215876 may include depth to the eye. Excitation of the smoke at each point of the axial scan of the shaft is known as "Z-Through" to determine the edge-to-beat position of the eye; and to measure the position of the eye to the position of interest; and to lie in the direction of the axial scan The light source is used to perform the flat connection of the eye, and the points of the flat drawing are combined with the class. Killing π ^ 劳 The light intensity and fluorescence of the condensed powder of the genus Time: When the substance is produced, it is found in the eye to indicate that the two sheep are bred. This method enables the amyloid protein such as the r-phase sputum-like powder-like condition. In the specific example, the method can further include Having excitation 2: illuminating the eye with light of a suitable wavelength corresponding to the fluorescent light of the compound powder of the dinosaur powder; and detecting the fluorescing of the eye for the combination of the amyloid-like powder in the eye Peak region of emission spectrum The appropriate wavelength of light in the domain. The powder-like mouth-of-mouth can be compound No. 11. The excitation spectrum can have a peak near the end nm. The wavelength of the eye is increased or decreased by about 2 〇 (4) in the peak of the excitation spectrum. And the emission spectrum may have a peak of about (10), and the light received by the eye is detected at a wavelength within the emission spectrum that is increased or decreased by about 2 〇. In another embodiment of the present invention, a method is provided for A device for detecting amyloid in a mammalian eye. The device comprises a light source configured to emit light that illuminates the eye and has at least one of a wavelength of light, a polarization of light, or a combination thereof, each of which is suitable for At least one genus-like compound is fluorescing in the starch-binding compound when bound to amyloid-like compounds, the starch-like binding compound has been introduced into the eye and specifically binds to an amyloid indicative of an amyloidogenic disorder; and optical a unit configured to receive light comprising fluorescence emitted by the illuminating eye and to measure fluorescence of at least glory produced by a starch-binding compound such as amyloid-like 201215876 Ratio, based on at least the measured time decay rate allows discrimination of binding to amyloid protein class = binding in the presence of compounds of the eye. / 敉 In other related specific examples, the optical unit can be configured to measure the time decay rate of at least one of the following: molecular rotor type starch humanization = substance; Congo red or Congo red street biological class powder binding compound ;^ s starch-binding compound; golden amine derivative-like powder-binding compound ^ gold ^ amine G or golden amine G derivative starch-binding compound; thio: thioflavin τ derivative-like powder-binding compound; and sulfur A compound such as a flavin s or a flavin S derivative starch-binding compound. The optical unit can measure: the intensity of crab light produced by binding to a powder-binding compound such as a powder-killing protein. The optical unit can be configured to determine the amount of bound starch bound to the amyloid based on at least the intensity and time decay rate. The optical unit can pass the phase energy & $ Π state to determine the average number of photons with a specific attenuation in a particular eye region. The light source can include a pulsed laser. The device can further include an optical scan configured to scan light from a source at various locations in the eye. The optical scan consists of a mirror mounted on the translation stage and a wiper containing a galvanometer mirror. The optical scanning unit can be arranged to illuminate at least one of the sampling eyes using a light source, the sampling comprising performing an if measurement of at least one of the at least one region of the at least one region or sampling the at least one region using the illumination of the light source. At least one of the locations, the different locations of the samples comprising illuminating the points, planes, or volumes in the at least one region. The optical scanning unit can be arranged to sample one eye at a different location in the article domain. In an example, the optical scanning unit can be arranged through 201215876 to perform a planar scan of the eye using a light source in a continuous plane extending along the depth to the vertical axis of the eyelid 17 and the month τ. The 兮 、 , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , , And at least one of an enhanced charge coupled device (ICCD); such as a sag light detector. In other related embodiments, the apparatus can further include a time-correlated single photon counting module that receives an electrical signal from the photodetector unit indicative of a photon count from the fluorescent light of the eye. The apparatus can include at least one processor module' configured to determine a time decay rate of camp light based on a photon count distribution that varies over time channel units. The optical unit can be configured to distinguish spontaneous fusion of a starch-binding compound bound to an amyloid-like protein with the dorsal hall's spontaneous fluorescence of the ocular tissue and other non-specific particles and unbound starch-like sputum. The cymbal optical unit can be configured to discern at least one of the presence and amount of one or more of the following: a starch-like binding compound; a starch-binding compound that binds to an amyloid-like protein; and an amyloid-like protein. The amyloid-like protein may comprise aggregates or pre-amyloid protein aggregates. For example, amyloid may comprise beta starch. The amyloidogenic disorder may comprise Alzheimer's disease. In other related embodiments, the optical unit can be configured to discern the presence of amyloid based at least on fluorescence detection. The optical unit can be configured to determine the rate of delivery of the starch-like compound to the eye based on the detected fluorescence, the spatial distribution of the starch-like binding compound delivered to the eye, and/or the starch-binding compound at the corneal interface of the eye. Concentration gradient. The optical unit can be configured to determine at least one of a spatial distribution and a temporal distribution of the compound-like starch in the aqueous humor based on the detected fluorescence. The optical unit can be configured to enhance the position of the ocular interface (such as the lens capsule of the eye) based on fluorescent signals caused by natural fluorescence emitted by tissue of the eye. The optical unit can be configured to determine the position of the nucleus of the eye based on the following: (1) the distance from the special sputum scraping structure (such as the lens capsule interface or the corneal interface of the eye) or the change in the (Η) detection intensity metric ( Slope). The optical unit can be configured to determine at least one dimension of the anatomical structure or substructure of the eye based on natural fluorescent excitation of at least a portion of the anatomical structure or substructure, wherein determining the at least one dimension can comprise at least 2 =: determining The thickness of the structure or substructure, the shape of the structure or substructure, and the distance between one or more structures or substructures of the eye. In other related embodiments, the optical unit can be configured to scan within the eye to determine the excited natural fluorescent light and thereby determine at least one region of interest in the eye; and use the light source to illuminate at least one region of interest of the sampling eye, The sampling includes performing at least one of the at least one complete region of the at least one region or at least one of the different locations within the at least one region using the source illumination to illuminate at least one of the regions, planes, or volumes in the region At least one of the samples; wherein the sampling is for determining the fluorescence intensity of the fluorescent light and the time decay rate of the fluorescence generated by at least one sampling region bound to the starch-binding compound such as amyloid. For example, the optical unit can be configured to determine the excitation natural fluorescence at various points along the depth of the eye into the scan (Ζ scan) and thereby determine at least one of the positions of interest of the eye; Fluorescent fluorescence generated by binding to a starch-binding compound such as amyloid at each point of each of 201215876 by using a light source to measure at least a plane in the direction perpendicular to the scan. Intensity and time decay rate of fluorescence. The device can be configured to enable immediate search for amyloids in the eye indicative of amyloidogenic conditions. In other related specific embodiments, the light source can be configured to emit light having an appropriate wavelength for a peak region of a fluorescent excitation spectrum of a starch-binding compound bound to an amyloid-like compound in the eye, and the optical unit can be It is configured to detect light having an appropriate wavelength for the peak region of the fluorescence emission spectrum of the fenestate-binding compound bound to the amyloid-like compound in the eye. The starch-like binding compound may be the No. 1 compound. The excitation spectrum can have a peak of about 47 〇 nm, the source is configured to emit light at or near the peak of the excitation spectrum, and the emission spectrum can have a peak of about 580 nm, and the optical unit name is configured to detect Add or subtract light within about 20 nm from the peak of the emission spectrum. Loxobin can be an indicator of amyloidogenic conditions. In another aspect of the present invention, there is provided a method for diagnosing or susceptibility to an amyloidogenic condition in a mammal 1 , such as a primate (such as a human), a dog, a drawing, and a material. , cattle and similar sputum: the method comprises illuminating a mammalian eye with a source having at least a wavelength characteristic, a polarization characteristic, or a combination thereof. The properties are each suitable for binding at least a type of starch binding compound to the indicator In the case of a silty protein such as an amyloid disorder, the compound A is produced in a fluorescent form, and the starch-like complex is not introduced into the eye and specifically binds to an amyloid which is indicative of an amyloidogenic condition; the reception includes irradiation Fluorescent light produced by the eye; and determining the rate of decay of at least the time of the starch-binding compound bound to the powder of the genus-like powder protein 12 201215876 2 =, which allows for at least time-based determination. In combination with the powder of the hall powder protein, it is present in the eye. The normal pair of starch-like compounds in the eye and amyloid-like proteins, Zhao Yushi in the South + Peru _ Autumn media, Guangguang: ""Daily showed in the mammals to cause starch to promote the development of money powder The risk of a condition. The halllet powder-like illness can be Alzheimer's disease. In another embodiment of the invention, a method of anatomical structure in an animal's eye is provided. The method comprises illuminating the eye with a source of at least one of a characteristic of 2, a polarization characteristic, or a combination thereof, each of which is adapted to produce a natural camp light in an anatomical structure of the eye; and a natural light produced by the light source in the eye The location where the intensity of the fluorescence changes the most, and the assay allows for an anatomical structure that varies most based on the intensity of natural fluorescence. In a particular and inscription, the device described herein is used in the method according to one of the specific examples of the present month. In other related specific examples, the anatomical structure can comprise an anatomical structure of the anterior segment of the eye. Identifying the anatomical structure can include determining the location of the anatomical interface based on determining where the natural camp intensity increases most, such as determining the location of the lens capsule interface of the eye. Identifying the anatomical structure can include determining at least the following natural glory measurements produced by the light source in the eye: corneal thickness of the eye, corneal shape, depth of the aqueous humor, lens shape, lens thickness, and lens substructure (eg, lens capsule, cortex, The thickness and/or shape of the core, the core; and may comprise determining the intraocular distance between at least two anatomical structures of the eye. The method can further comprise detecting a powder protein indicative of an amyloidogenic condition in the eye of the mammal using a light source. The method may include 13 201215876 comprising irradiating a mammalian eye with a light source, the light source further comprising at least one of a wavelength characteristic, a polarization characteristic, or a combination thereof, each of the characteristics being suitable for binding at least one starch-like binding compound to the indicator starch In the case of a powdery protein such as a disease, a fluorescent compound is produced in the starch-binding compound, and the compound is incorporated into the eye and specifically binds to an amyloid-like disorder; Fluorescent light produced by the eye; and measuring the time decay rate of at least the fluorescence of the fluorescence produced by the starch-binding compound bound to the amyloid-like protein, the assay allowing discrimination based on at least the time decay rate to bind to an amyloid-like protein or the like The presence of a starch binding compound in the eye. Identifying the presence of a compound binding protein-like compound bound to an amyloid-like protein may comprise spontaneously fluorescently binding to a starch-binding compound such as an amyloid-like protein to the background of the ocular tissue and other non-specific particles and unbound starch-like compounds. Light. This method enables the immediate search for amyloids in the eye indicative of amyloidogenic conditions. The method may further comprise illuminating the eye with light having an appropriate wavelength for a peak region of a fluorescent excitation spectrum of a starch-based sounding compound that binds to an amyloid-like compound in the eye; and detecting that the eye receives a combination with the eye in the eye Starch such as powder protein = light of a suitable wavelength in the peak region of the fluorescent emission spectrum of the compound. The starch-like binding compound may be the compound No. 11. The excitation spectrum may have a peak of about 470. The eye is illuminated at a wavelength of the excitation spectrum plus or minus a wavelength within about 20 nm' and the emission spectrum may have a peak of about 580 nm to detect the peak of the emitted light of the eye at the emission spectrum. Add or subtract at a wavelength of about 2 〇 nm. ~ In other handle specific examples, a method may allow at least two different firefly 14 201215876 light clusters having similar fluorescence spectra in the eye to be distinguished based on at least the time decay rate, the similar fluorescence spectrum comprising the emission spectrum and the excitation spectrum At least one of the significant overlaps. A method may further comprise a knife cloth that exhibits at least one of a fluorescence intensity and a life decay of at least one of the fluorophores in two dimensions. Further, the method may include glory intensity and lifetime decay based on at least one fluorophore At least _ the number of photons combined in the eye and the number of unbound photons. One method may comprise the distribution of glory intensity and lifetime decay of a compound of the genus of the prion protein and the compound of the dinosaur powder which is not bound to the protein in two sizes. In two sizes: now comparable to at least one of the scanner and the laser. The method can further be determined by averaging the intensity of the camping light associated with the decay of the lifetime over a particular area of the eye. parameter. Additionally, the method can further include aligning the alignment source with the eye along the confocal path to determine a reference point in the eye. In another embodiment of the invention, a method for determining a glomerized group bound to an eye = protein is provided. The method comprises irradiating an eye group with a light source having at least one of wave I: stem polarization characteristics or a combination thereof, each of which is adapted to be suitable for binding to at least one starch-like binding compound in the class of powder-binding compounds Produces fluorescence, such a starch knot "Do not have been introduced into the ocular tissue and specifically bind to the protein; receive the package by:, the % of the light produced by the eye; and the determination of at least by binding to the protein: The time decay rate of the luminescence of the glare produced by the powder-binding compound, = based on at least the time decay rate, distinguishes the presence of the unbound, sputum & compound in the ocular tissue. As described in the accompanying drawings The foregoing description will be apparent from the following detailed description of the embodiments of the invention, the same reference Specific Embodiments [Embodiment] Specific examples of the present invention are described below. According to an embodiment of the present invention, a non-invasive, early and reliable detection of a starch-like egg is provided. Systems and methods for raising or forming aggregates. In some embodiments, the detection of powders and/or aggregates is indicative of an amyloidogenic condition. AD, familial sputum), sporadic AD, Crentzfeld-Jakob diSease, variant gram-Asian's disease, spongiform encephalopathy, prion diseases (including scrapie, bovine spongiform encephalopathy) And other veterinary virulence diseases (Pri〇nopathy), Parkinson's disease (Panins (10) $ disease), Huntingt〇n (sdisease) (and trinucleotide repeat disease), muscular atrophy Lateral cord sclerosis, D〇wnis Syndrome (Dijon trisomy (d) (D) (D), Pick's Disease (frontotemporal dementia), Body disease ), neurodegeneration associated with brain iron accumulation (Hallerv〇rden-Spatz Disease), synuclearopathy (including Parkinson's disease, multiple system atrophy, Lewy body dementia and other diseases), nerve Inclusion body disease 'tau protein (tauopathy) (including progressive nucleus, Pick's disease, basal ganglia degeneration, hereditary frontotemporal dementia (with or without Parkinson's disease), pre-onset neurodegenerative state, and Guam muscle Atrophic lateral sclerosis (Guam amy〇tr〇phic WeM scler〇Sis) / Parkinson's disease - dementia syndrome) β These conditions can occur alone or in combination with 16 201215876. Amyloid analysis is also suitable for detection Infectious spongiform encephalopathies (TSE), a prion-mediated disease characterized by degenerative cavernous nerve degeneration in the brain and associated with severe and fatal neuropathy and symptoms. TSE prion diseases include gram-Asian disease (cjD); new variant gram-Asian disease (nv-CJD); Gertzmann-Straussler-Scheinker syndrome Fatal familial insomnia; Kulu disease; KUru; Alpers Syndr〇me; Bovine spongiform encephalopathy (BSE); scrapie; and chronic wasting disease > CWD). Diagnostic methods can be performed on mammalian eye tissue, such as primates (4) (such as humans), dogs, cats, stocks, cattle, and the like. The individual to be tested (e.g., a human subject) includes individuals (patients) suspected of having such conditions "individuals at risk of developing such conditions. For example, g, individuals with a family history of AD or other risk factors* (such as advanced age) are tested using the techniques described herein. Individuals who are not known to be at risk of developing or developing such conditions are also tested. The diagnostic method is carried out by contacting an eye group of a mammal (for example, a human individual) with a protein associated with a class of powder (for example, a P-class powder (A(3)) t campon group." (amy丨.id prQiein)" actually said: = neuritis dementia plaque (four) protein f or peptide, regardless of the aggregation of protein powder (70 king or part). Amyloid protein is preferably starch-like precursor egg: ( APP) or App (for example, naturally occurring) proteolytic cleavage, such as Αβ. APP cleavage products include Api 4〇, A (32 4〇m and oxidized or paternal Αβ. Fluorescent compounds can also be combined with natural There are APP and Αβ variants of 201215876, including single nucleotide polymorph (SNP) variants. Fluorescent compounds can (but are not required) bound to beta starch aggregates. Binding to beta starch aggregates A discussion of fluorophores can be found in G〇ldstein et al., "Cytosolic beta-amyloid deposition and supranuclear cataracts in lenses from people with Alzheimer's disease,"

Lancet 2003; 361: 125 8-65, the entire disclosure of which is incorporated herein by reference. Aggregates containing Αβ (a pathogenic protein accumulated in AD) have been found to form a nuclear/deep cortical cataract in the lens and brain of patients with Alzheimer's disease. Aβ deposits were collected as intracellular aggregates in the cytosol of lens cortical fiber cells. It has been shown that lens A (3玖 is present in the adult lens in the same amount as the soluble apparent monomer and dimeric substance in the normal adult brain. The substantial part of the lens Αβ binds to other lens proteins, including sufficient lens structure protein αΒ Crystalline globulin. (6) Crystalline globulin exhibits intermolecular binding affinity in the in vitro tube and is co-immunoprecipitated from tannic acid-treated human lens tissue homogenate, indicating a strong protein-protein association. Human Αρι_42 with ρ Increased content promotes lens protein aggregation. Blocking 蚪-enhanced lens protein aggregation by metal chelation or reactive oxygen species scavengers, indicating that metalloprotein redox reaction involves the lens protein aggregation process and cataract formation of the AD towel The data showed that there was a pathological phase between the Αβ and the lens protein. In addition, the 郸-mediated reaction in the lens indicates that the sputum powder-like substance is particularly involved in the human cans of AD pathophysiology_42 18 201215876 质 'To promote lens protein aggregation and nuclear / epithelial An effective oxidative peptide for the formation of cataracts. Further information regarding protein aggregation and cataract formation can be found in U.S. Patent No. 7,1,7, 〇92, to the name of the entire disclosure of which is incorporated herein by reference. In one embodiment of the invention, an increase in the binding of a fluorophore compound to an ocular tissue (e.g., an intracellular compartment of a lens cell) compared to a normal control indicates that the mammal is suffering from AD or is at risk of developing AD. As used, "fhl〇r〇ph〇re"j or "fluorophore compound" is any substance that has the desired fluorescent characteristics when irradiated with light of a specific wavelength and/or polarization characteristics. Preferably, in this context, H彳# t ' Μ ® is a "starch-like binding compound (such as ~ compound )" which, as used herein, means a compound that binds to an amyloid protein, wherein "amylidoid prion" (amyl〇idpr〇) Tein)j as defined above. The fluorophore may be a starch-like binding compound that lacks fluorescence when exposed to light of a particular wavelength and/or polarization characteristic. Additionally or otherwise ': :营: The combination of the part of the standard iron and the compound of the powder-like compound:: , where the part of the compound of the powder of the class is not present in the iron of the cursor; it will not show the characteristics of the desired fluorescent. In any medium using fluorophores, two =: can be different from the time of uncombined. Can be different from the light intensity of the luminescence of the camping group. It can be changed when it is not combined. The compound of the No. u (the lower coat ratio is in the step-by-step discussion) "When the compound is combined with the text: Figure 5

S 19 201215876 Fluorescent clusters that change when not combined. Further discussion of these properties of fluorophores (especially compound No. 11) can be found in J. Sutharsan et al., "Rational Design of Amyloid Binding Agents Based on the Molecular Rotor Motif", ChemMedChem 2010, 5, 56-60, The entire disclosure is hereby incorporated by reference. Preferably, the camping light compound is bound to another segment of Αβ 1 -42 or a powder-like precursor protein (App). The fluorophore compound preferentially binds to amyloid-like protein compared to proteins containing other β-pleated laminates. As noted above, the fluorophore compound can contain a fluorescent probe or can be used as a fluorophore without the addition of a fluorescent probe. For example, the fluorescent probe or fluorophore can be a golden amine or a golden amine derivative compound such as {(anti, trans), 1-bromo-2,5-bis-(3-hydroxycarbonyl_4_ Hydroxy) cinnabaryl benzene (BSB) }. In a particular embodiment, the fluorophore can be Compound No. 11 (discussed further below in connection with Figure 5), which is a fluorescent compound designed according to molecular rotor motifs. According to an embodiment of the present invention, the powder-like compound may be a molecular rotor, a golden amine, and/or a golden amine derivative. An exemplary fluorophore is described in U.S. Patent No. 4,249, the entire disclosure of which is incorporated herein by reference in its entirety in its entirety in the the the the the the the the the the the the the the -(3_Pentyl-4-yl-based) cinnamylbenzene (Special: Golden amine G and its derivatives are known in the art of the if (for example, US ▲ 3'259; No. 6,168,776; 6,114,175). Use fluorescein Αβ peptide 'but not light-emitting. Diagnostic method can also use it: golden amine guanidine derivative to detect scorpion peptide in the eye. Probes. These fluorophores and probes market# are sold, for example, from Mn1, 1 Τ% with ecular Pr〇bes, Eugene, OR,]js Α. 20 201215876

Some dyes (eg X-34 or {(and, and), _ι·bromo-2,5_bis-(3-hydroxyl-4-yl) cinnamylbenzene (BSB) }) (Styren et al, 2000, J

Histochem. 48: 1223-1232 (Link et al., 2001, Neurobiol. Aging 22: 217-226; and Skrovonsksy et al., 2000, Proc. Natl. Acad.

Sci·U.S.A. 97:7609-7614) has been used to analyze brain tissue (not eye tissue). These probes emit light in the blue and green range, so the diagnostically relevant fluorescence content exceeds the amount of spontaneous fluorescence of the human lens in the blue-green range. Other suitable compounds include detectable methoxy reagents such as Me_x〇4 [1,4-bis(4'-hydroxycincosyl)-2-methoxybenzene> Other methoxy reagents include, for example, golden amines or golden amines. Derivative compounds such as {(and, and bromo-2,5-bis-(3-carbylcarbonyl-4-hydroxy) cinnamylbenzene (BSB) oxime. These compounds are described in Mathis et al, Curr pharm _, U.S. Patent No. 1, 316, pp. The method can be used in this article. Other types of powder-binding probes can also be used, such as θ-T, thioflavin s, Congo red dyes, the aforementioned derivatives, or other bamboo organisms. Further information on the compounds can be found in U.S. Patent No. 7,297,326, the entire disclosure of which is incorporated herein by reference. Patent Shenyue: No. 2GG9/0G41666 and US Patent No. 7^07,092 Shen patents and eye pattern entire teachings are incorporated herein by reference in a specific embodiment, ϋ an Μ 11 M M c τ discussed further in connection with FIG. 5). 21 201215876 Additional information on related methods, amyloid-like disorders, amyloid-like and fluorophore compounds can be found in U.S. Patent No. 7,297,326, to Goldstein et al., U.S. Patent No. 7,1,7, to Goldstein et al. 〇92 and

No. 6,849,249 to Goldstein et al., the entire disclosure of which is incorporated herein by reference. In addition, the above-mentioned other teachings afl can be found in U.S. Patent Application Publication No. 2,8/88,875, U.S. Patent Application Publication No. 2009/0041666, the entire disclosure of which is incorporated by reference. Into this article. The methods provided herein can further comprise comparing the lens fluorescence of the test patient after administration of the fluorophore to a suitable control group. Examples of suitable control agents include the fluorescent content of endogenous spontaneous fluorescent or non-AD individuals (or non-AD individual populations) of non-AD individuals (or individual populations) after administration of the fluorophore. According to one embodiment of the invention, the amount of amyloid present in the eye found based on the techniques disclosed herein can be compared to a statistical analysis of the amount of amyloid-like protein indicative of a risk of a disease condition or a development of a disease condition. Without wishing to be bound by theory, it is desirable for a healthy adult to have at least some minimal amount of amyloid in the suprarenal region of the lens of the eye. The techniques disclosed herein can therefore be used to determine if an individual has an amount of amyloid in the eye that is above the statistically significant amount of normal control amyloid content in the eye. A study of the deposition of amphotericin in the eyes of individuals with Alzheimer's disease can be found in Goldstein et al., "Cytosolic beta-amyloid deposition and supranuclear cataracts in lenses from people Whh Alzheimer's disease," Lancet 2003; 361 ·· 125 In 8-65, the entire disclosures of which are incorporated herein by reference. 22 201215876 As discussed below in conjunction with the actual implementation of the invention, it has been shown in the -Characteristics of the present invention that it is comparable to uncombined powder-binding compounds, ... between the class of starch-binding compounds Identify. Specifically, the results in Test 1 have found that the time decay rate of the unbound fluorescent starch-like compound (No. 11 compound) herein is, for example, μ nanosecond: and the starch-binding compound as described herein is bound to the starch-like compound. The time decay rate of the protein (aggregated starch (Αβ) peptide) is, for example, 2 25 nanoseconds. According to an embodiment of the present invention, the detection of the unbound starch-like binding compound (the compound No. 11) can be expressed as a time decay rate of 14 nanoseconds plus or minus 3 seconds, and the bound starch-like starch is bound. Detection of the compound of compound u can be expressed as a time decay rate of 2.25 nanoseconds plus or minus 0.3 nanoseconds. Other decay rates and confidence levels for the differential starch-like compounds and amyloid-like proteins can be used. A method and apparatus for detecting fluorescence of a beta-like starch-labeled beta-starch (Αβ) protein in an eye lens according to an embodiment of the present invention, and use thereof. In one aspect, the apparatus provided herein is an optical imaging device that employs a fluorescence scanning mechanism in combination with a lifetime spectrum to enable detection of fluorescent molecules and provide information about their spatial distribution and their surrounding environmental characteristics. According to one embodiment of the invention, the device (eg, a multi-function optical scanning and fluorescent system) enables recognition of the anatomy of the anterior segment of the eye based on natural fluorescence excitation; and provides spatial information about the anterior segment of the eye (such as corneal thickness and The lens shape) and provides an intraocular distance. In addition, the multifunctional optical scanning system 23 201215876 according to an embodiment of the present invention provides an in vivo ocular pharmacokinetic study for an exogenous fluorescent-like powder-binding compound that is not bound to amyloid in the eye. tool. For example, the system measures the gradient intensity of a class of powder-binding compounds at the corneal interface (such as the tear gland/corneal epithelial interface). In addition, the system measures spatial and temporal information about the bioavailability of the powder-like compound in the aqueous humor. Further, the multifunctional optical scanning system of one embodiment of the present invention allows detection of fluorescent molecules and differences therebetween based on optical properties such as fluorescence decay time (τ). The system allows the detection of labeled fluorescent quinone-like conjugated compounds bound to Αρ in the lens of the eye; detection of natural fluorescence in the eye; and identification of (1) labeled fluorescent starch-binding compounds bound to Αβ in the lens of the eye and (ii) Natural fluorescence in the eye. As used herein, "natural fluorescence" means natural fluorescence in the eye that can occur independently of the introduced developer. 1 is a schematic view of an optical device according to an embodiment of the present invention. Fluorescence excitation is achieved by a pulsed laser beam with a numerical aperture lens 1 〇 1 focused in the eye. The time-dependent single photon counting (TCSPC) technique is used to construct two confocal configurations with a confocal configuration of the fast spur photodiode detector (APD) 102. The TCSPC is performed by repeatedly exciting the sample (eye) 103 with light of a short pulse and recording the subsequent fluorescence emission that changes with time. This operation is generally performed within a 2 nanosecond time scale. Also in the specific example of Fig. 1, the identification of the anatomical structure of the lens is performed by scanning the objective lens 1〇1 on the shaft using the translation stage 104. The signal at each point along the scan is measured to reveal the anatomical structure of the anterior segment (such as the cornea of the lens, 24 201215876 lens capsule and supra-nuclear region). In addition, scanning: applied to the pharmacokinetics of exogenous starch-binding compounds not only provides the airborne powder-binding compound, but also provides a concentration that penetrates the cornea. The vaginal cornea enters the starch-binding compound such as the eye liquid. In the specific example of the figure, once the natural camping light is known to be the position of interest in the eye, it is in the 'and electric' optical of the ten mirror 105. The vertical plane of the axis (xy): tracing. To ensure that the fluorescence decay of the measurement is assigned to the corresponding position of the two-dimensional scan. The text 'synchronizes the galvanometer group scan with the pulse and green measurement of the time-dependent individual photon count. Xy# describes the glory decay time information of each point of the image. In the specific example of Figure 1, you can use the special hardware module and / or use a specific stylization to implement the module = computer execution - or multiple The module includes, for example, a frame receiver module, a group, a τ calculation module, and a scanner control module. The general purpose computer and/or the bee is receiving data from each other. In the specific example of FIG. 1, for the time-dependent individual photon counting, the decay curve of the spontaneous camping light at each scanning position of the lens is recorded and thus the two-dimensional representation of the fluorophore distribution can be evaluated and analyzed based on the glory decay time and intensity. The image of the calculated decay time can be encoded by pseudo-color and the disc-intensity image can be overlapped for better clinical interpretation. Since the fluorescence decay time is characteristic of nine molecules per two battalions, we can measure and separate the excitation in the sample volume. Camp Light Group (from the crystal cans of the cans of the genus 25 201215876). By combining the fluorescence intensity and lifetime metrics, additional size information is obtained to distinguish between several fluorescent markers. As discussed herein, one of the present inventions A device of a specific example can include a light source. As used herein, a "light source" can be any light source that can be configured to emit light that illuminates the eye and has at least one of wavelength and polarization characteristics of light, wherein light The wavelength and polarization characteristics are such that when at least one of the dendrimer-binding compounds binds to the amyloid-like protein, the starch-binding compound produces fluorescence in such a manner that the time decay rate of the fluorescent light can be determined based on the glory received by the irradiation. In one embodiment of the invention, the light source can be configured to emit light having an appropriate wavelength for a peak region of the luminescence excitation spectrum of the starch-binding compound bound to the amyloid-like compound in the eye, and the optical unit It can be configured to detect light having an appropriate wavelength for the peak region of the fluorescent emission spectrum of the fenestate-binding compound bound to the amyloid-like compound in the eye. For example, when the starch-like binding compound is compound No. 11, the excitation spectrum has a peak of about 470 nm, and the light source can be configured to emit a peak of about 47 〇 nm plus or minus about 20 nm, such as 470 nm plus or minus 5 nm, plus or minus 10 nm, plus or minus 15 nm or plus or minus 2 〇 nm. In addition, the emission spectrum of Compound No. 11 has a peak of about 580 nm, and the optical unit can be configured to detect a peak at about 580 nm plus or minus about 2 〇 nm, such as 580 nm plus or minus 5 nm. Internal, plus or minus within 1 〇 nm, plus or minus 15 nm or plus or minus 20 nm. In general, there is typically a displacement between the peak of the excitation spectrum of the fluorescent compound and the peak of the emission spectrum. According to a specific example of the present invention, it is suitable to use a compound whose emission spectrum peak is shifted relative to the excitation spectrum to enable differentiation of the natural spontaneous crab light of the fluorescing eye of the combined fluorophore. For example, an emission spectrum with a peak greater than about 5 Å is advantageous for distinguishing from natural spontaneous fluorescence of the eye. It has been confirmed that the ninth compound is suitable for this purpose, and has an emission spectrum having a peak of about 580 nm and a significant shift in an excitation spectrum having a peak of about 470 run. Fig. 12 is an emission spectrum of a fluorescent-type starch-binding compound (the ninth compound) excited at nm in an embodiment of the present invention. Those skilled in the art will readily appreciate other excitation and emission spectra that can be used based on the above. According to one embodiment of the invention, the apparatus may use an "optical unit", as used herein, to mean that it is configurable to receive light comprising fluorescence generated by illuminating the eye and to determine at least by combining Any single A' formulation of the fluorescence decay time rate of fluorescence produced by a starch-binding compound such as an amyloid allows discrimination based on the temporal decay rate to distinguish the presence of a starch-binding compound such as an amyloid-like compound in the eye. For example, referring to FIG. 1, the optical unit may include one or more of the following: the objective lens 10, the translation stage 104, the scanner 105, the photodetector 1, the camera, the LED, the plurality of lenses, the aperture, and the beam splitting. , dichroic filter, time decay calculation, group, frame receiver module, TCSPC module and scanner control module. Optics: Some of the functions of the 可 can be performed by a specific stylized general purpose computer or dedicated hardware, for example for performing time decay calculations. According to an embodiment of the invention, the function of the objective lens 1 (H, the translation stage 1〇4 and the scan || having the current mirror 1G5 can be performed using a plurality of different possible devices instead of or in addition to the components. In general, the translation stage The function of the objective lens and the scanner with the galvanometer mirror is referred to herein as "optical sweep 27 201215876 7 unit s_ing unit", which can refer to any device J that performs the equal function of the scanning beam on the desired region. , said, including the purpose of measuring the reference point in the eye and the eye ==. The optical scanning unit can perform a function of inducing movement of the lens = movement of the lens or the multi-dimensional path of the lens along the movement; and knowing the region of the region on the region of interest, for example, performing the beam in the region of interest - A volume or other type of scan, such as in a mirror or other optical device, induces motion in the optical path of the beam. In the specific example of Fig. 2, the use of a confocal arrangement means that there is no need to use expansion = two, for example, in a system that requires light to enter the off-axis of the eye at, for example, a 45 degree angle. This is convenient for the patient. Figure VIII is a graph showing the natural fluorescence intensity versus displacement measured during the algorithmic period of the lens interface for scanning the illumination path along the eye (Ζ scan) and Figure 2Β A specific example of the present invention is a 13⁄4 derivative diagram of Figure 2A. The basic principle of the algorithm is the natural fluorescence intensity value. The position of the force port is the reasonable indication that the maximum per unit scan distance is the opening of the lens boundary. For + + s, the algorithm measures the distance from the starting point of the z撼松 known to the maximum inflection point of the fluorescence intensity. In a specific example, the algorithm is performed as follows, and can be operated immediately: / Collecting data in a two-dimensional array, wherein the first (self-variable) dimension is 疋, the distance from the starting point for the code thief measurement, that is, the scanning distance, And the first dimension (variable) is the glory intensity measured via a photon detector (APD). 28 1) Use the 5-point moving average type to rotate the data array to smooth the strong 201215876 degree', ie remove the high-frequency noise that interferes with the differential. 3) Use the differential type to rotate the smoothed data array to obtain the first derivative of the intensity column. 4) Search for the maximum derivative intensity value t-intensity first-order derivative array. This is the biggest recurve point. Determine the corresponding scanning distance. As shown in Fig. A and Fig. 2B, the lens capsule can be measured using the above technique. In addition, the position of the anatomical structure (such as the cornea, the aqueous humor) and the distance between them can also be determined. A delta can be applied to specify the measured distance along the reference plane of any axis. Fig. 3A and Fig. 3B are diagrams for explaining the measurement of the fluorescence agronomic time according to an embodiment of the present invention. The fluorescence decay time can be calculated by fitting the index (Fig. from) and the intensity curve (herein, in photons/second) with respect to time (here, in nanoseconds). It can also be obtained by linear fitting with the slope (referred to as "the decay rate 〇f fluorescence" as used herein to indicate the characteristic time constant of the decay curve of the fluorescence intensity; for example, the exponential time f The slope of the number or the curve of the fluorescence decay curve. The above algorithm of the port diagram 2Α, Fig. 2Β and Fig. 3a, Fig. 3Β can be performed, for example, using a special special hardware module and/or a general purpose computer using a specific stylization to execute the above algorithm. The modules may, for example, use or receive data from a TCSPC module, a frame receiver module, and a τ calculation module of a specific example of the figure. Figure 4 is a time related list illustrating a specific example in accordance with the present invention. Schematic diagram of the use of photon counting. The pulsed light source 4〇6 repeatedly excites the sample. The detector unit smashes the photodetector (APD) and the sample is detected by the sync module (SYNC) 407. The excitation flash (CFD) 408 only responds to the first photon detected by the detector 402, independent of its amplitude. The first photon from the sample emission is a time-amplitude converter (TAC). The stop signal β excitation pulse trigger start signal of 409. The multi-channel analyzer (MCA) 4 1 0 records the repeated start/stop signal of the single photon event of TAC 409, and generates a photon count histogram of the channel unit change with time. This histogram calculates the lifetime. The MCA can be implemented using dedicated special hardware modules and/or general purpose computer programs that use specific programming to perform such tasks; and can be communicated with a particular stylized general purpose computer data. In the examples, a system comprising a fluorescent starch-binding compound and device is intended to aid in the diagnosis of a patient's possible Alzheimer's disease after appropriate clinical examination. These patients have symptoms or signs consistent with Alzheimer's type dementia. The device employs a confocal scanning mechanism in combination with fluorescence lifetime spectroscopy techniques that enables the identification of the smear-like structure of the anterior segment of the eye and the identification of fluorescent fluorophores based on their optical characteristics. Figure 5 shows Compound No. 11. a structure which can be used as a fluorescent starch-based binding compound as a specific example of the present invention. A fluorescent compound designed and shown to bind to aggregated p-type starch (Αβ) peptides. This is combined with natural fluorescence, indicating that the quinone compound is found in the lens tissue of patients with Alzheimer's disease. A good candidate for in vivo markers of Ap aggregates. The chemical name of the compound No. [ is [(E)-2-(2-(2-decyloxyethoxy)ethoxy)ethyl·2 • Cyano _3_(6_(piperid-1 -yl)naphthalen-2-yl) acrylate]. Further information on the quinone compound can be found in J. Sutharsan et al., "Rational Design of Amyloid 30 201215876

Binding Agents Based on the Molecular Rotor Motif j , ChemMedChem 2010, 5, 56-60, the entire disclosure of which is incorporated herein by reference. Compound No. 11 has been formulated into an ophthalmic ointment (No. u compound ophthalmic ointment) which contains about 5 mg/g of the i-th compound, 8 % of petrolatum and 20% of mineral oil. According to one embodiment of the invention, the fluorophore-type starch-binding compound can be applied to the eye of an individual to be tested in any of a number of different possible forms. For example, the fluorophore-type starch-binding compound can be administered in the form of an ointment, in the form of a solution, using a contact lens, by injection, in liquid form, in solid form, by iontophoresis or by other techniques. The apparatus of one embodiment of the present invention is designed to detect fluorescence in a confocal detection procedure in a time domain with high sensitivity and speed. The device has two main functions: 丨) using a translation stage and galvanometer scanner to transmit and scan the beam of light reaching the front of the eye (such as the nucleus of the lens) and 2) identifying and discriminating fluorescence based on fluorescence lifetime measurements Sexual fluorescent group. The device of the present invention-specific example uses an axial scan 4 Ζ scan eye anatomy structure based on laser excitation of the ocular tissue along the eye field 、V, the axis of the day, to obtain the intraocular distance Police ~ Beixun. The ζ scan reveals a plot of natural fluorescence intensity as a function of depth, & provides information about where to wait; The location of the target can be exemplified by the 7 7 7 U, the nucleus of the 頬 φ Β 。. Scanning can be done in a few seconds, for example 2 曰曰^, within 1 2 seconds, such as about 0.2 seconds, 0. 3 seconds, 〇.4 seconds, 〇·5 seconds, 〇6 υ·7 seconds, 〇β seconds, 〇·9 seconds, iO seconds, 12 seconds, 14 seconds, 丨6 · 8 31 1 · 8 seconds or 9 η 2 seconds to reduce eye movement artifacts, or in 201215876 2nd time suitable for reducing eye movement artifacts Completed within the volume. Additionally or alternatively, the device can be tracked with eye movements - used to reduce motion artifacts. Piezoelectric drives, linear motors and other controls can be used for this (4). Axial scanning may also allow for the measurement of the bioavailability of starch-like, di-compounds at the ocular interface (such as the tear gland/corneal epithelial interface) and the starch-like binding compound in the aqueous humor. The apparatus of the present invention, which is a specific example, recognizes the fluorescent knife by performing Xy scanning. The sample of the towel is raster scanned by the device driven by the electric material. The fluorescence lifetime of each scan position of the human scorpion is recorded, and thus the two-dimensional representation of the fluorophore distribution can be evaluated and analyzed based on the glory decay rate and intensity. A two-dimensional table of glory distribution based on decay lifetime... (but not necessarily) includes a two-dimensional representation based on fluorescence intensity', referred to herein as "fluorescence lifetime image." According to one embodiment of the invention, the fluorescence lifetime measurement is based on repeatedly exciting the eye with a short laser pulse and recording subsequent glory emission as a function of time. Since the fluorescence decay time is characteristic of each fluorescent molecule, it is possible to measure and separate the powder-like compound from the lens (4) light excited in the sample volume. In particular, in one embodiment of the invention, the fluorescence lifetime measurement can be obtained by time-dependent early photon counting (TCSPC). Scanning speed and data acquisition can be synchronized and executed, for example, within 0.5 seconds to reduce any eye movement artifacts, or another amount of time suitable for reducing eye motion artifacts. The TCSPC principle is based on the detection of single photons emitted by a pulsed laser and the detection time of individual photons arriving. When photons are detected, the time of the corresponding detector pulse is measured. Events for many detected photons are collected in 2 32 201215876 memory. The fluorescence decay lifetime can be calculated by constructing a histogram from an individual time measurement. The apparatus of one embodiment of the present invention operating in the TCSPC mode can achieve, for example, a count rate of about 1 〇 7 photons per second. Therefore, 1 〇 4 photons can be collected in less than 1 millisecond. These counting rates are important when high speeds are required to obtain fast scan information for the human lens of the moon. Other count rates are available. The device of one embodiment of the present invention can be designed to obtain specific information from a particular location of the human eye lens. Examples of such locations include the nucleus, lens capsule, nucleus, cornea, and aqueous humor. This is achieved by precisely aligning the individual, understanding the anatomy of the eye, and obtaining the fluorophore k 中 in the scanned area of the lens with high specificity and sensitivity. BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a schematic illustration of an optical platform (also discussed above) of one embodiment of the invention. Fluorescence excitation is achieved by a pulsed laser beam focused in the eye 103 by a high numerical aperture objective lens 101. Although other repetition frequencies and pulse widths may be used, the laser may pulsate, for example, at a repetition rate of about 40 Μ Η z and produce a pulse of about 200 picoseconds wide. For example, a repetition rate as low as about 1 MHz to about 240 可使 can be used, and a pulse width of about 4 皮 picoseconds to about 400 picoseconds can be used. The beam is then reflected off the pair of galvanometer scanners and focused by a high value objective attached to the translation stage 1〇4. Determine by first aligning a blinking eye with the device and performing a η Ζ scan

The position of the region of interest 6 ρ η Τ, Θ 9, ν 4* U 尺心直IROI) and 2) Xy scan to obtain specific information on the area within the nucleus to obtain fluorescence measurement on the nucleus of the eye . According to a particular embodiment of the invention, the individual alignment consists of a focal plane that identifies the objective lens as a reference starting point for the measurement. Also used as a fixed target 33 201215876 Light diode (LED) is ring-shaped by the objective lens 101 at the corner of the eye 1〇3. Use the camera to visually reflect the ring reflection away from the corneal surface. Once this is done, an eye scan can be performed to get the necessary information. According to a specific embodiment of the invention, the use of the translational stage 104 optical axis (axis phantom scanning objective 1G1 to perform lens anatomy y involves exciting the natural light with a laser source and identifying the anterior segment: the membrane, the lens capsule and the lens The anatomical structure and basin phase of the nuclear region: two large separations. In addition, the scan provides information on the pharmacokinetics of the source starch, sigma 5 compound administered to the eye. In accordance with an embodiment of the present invention, the region of interest 4 in the eye is identified using a helium scan measurement, i.e., a planar scan (xy scan) is performed in a plane perpendicular to the axial sweeping cat using a galvanometer mirror. In order to ensure that the measured camping light sorrow curve is assigned to the corresponding position of the two-dimensional scan, the data acquisition plate of the galvanometer mirror T(10) is synchronized. A smart scan may require, for example, scanning a region of 50 x 5 〇 _ on the nucleus of the human eye within 5 seconds and finding the life sorrow value. It should be understood that 'are available in other sizes and locations, and scan time. In accordance with a specific embodiment of the present invention, detection is achieved using TCSpc by a confocal configuration with a fast burst photodiode detector (APD) 1〇2. The same objective lens 101 is used to collect the camping light from the excitation molecules in the form of an excitation laser. The two-way chromo-optic filter with other band-passing crests that suppress the residual scattered laser light passes through a small aperture to enable confocal detection. Using the fast-timing option, APIM 〇2 can, for example, provide a timing of photonic detection efficiency of 49% at 550 nm over a full-width of 5 〇 picoseconds (Fuu (Half)). 34 201215876 Resolution 'but other timing analysis can be used And photon detection efficiency. In accordance with an embodiment of the present invention, a description of data acquisition and electronics after TCSPC is shown in Figure 4 (also discussed above). The pulsed light source 4〇6 repeatedly excites the sample 403 at a repetition rate of, for example, 40 MHz, while the excitation pulse is detected by the SYNC module 407, which is also set to, for example, 40 MHz. The excitation pulse triggers the start signal. The constant ratio discriminator (8) only responds to the first photon detected by the detector 402, independent of its amplitude. The first photon from the sample emission is the stop signal of the time-amplitude converter (TAC) 4〇9. When the APD 402 detects photons, a short pulse is formed at the PMT output. The pulse is "cleared" by CFD 408 and entered into TAC 409 as a "stop" pulse. Once the stop pulse (i.e., the first arrival photon) is detected, the voltage ramp is stopped and the voltage value (equal to the time difference between the start pulse and the stop pulse) is transmitted to the multi-channel analyzer (MC A ) 4丨〇. McA 4 records the repetition start of the single photon event from TAC4G9. The stop signal is incremented and the count corresponding to the detection power (time) in the channel is increased. This process is repeated for each pulse, and finally after multiple cycles, a histogram of the number of photon changes over time channel elements is generated. This histogram represents the fluorescence intensity as a function of time, thereby obtaining a fluorescence decay lifetime. According to one embodiment of the present invention, rCiSTC (10) called with a time-decomposed mode of special time stamping can be used.

The GmbH' Bedin, Germany) Pc board performs data acquisition, which stores all relevant information for each photon detected for further data analysis. In detail, the arrival time of each photon is recorded in the detector in synchronization with the laser excitation pulse and the sample position and the number of detection channels. The acquisition board synchronization rate can be set, for example, to 35 201215876 for 40 MHz' time resolution of 4 picoseconds and channel count depth to epoch (other synchronization rates, time resolution, and channel count depth can be used). In accordance with an embodiment of the present invention, software acquisitions that can be performed using a P1C〇Quant (GmbH, Berlin, Germany) can be controlled via a tcp/IP network and synchronized with the galvanometer scanner and acquisition board via a TTL signal to define a map. Like the line and the box. For example, it can be recorded in the 尬 command mode and exhibits fluorescence lifetime and intensity images. "According to one (4) of the present invention, the fluorescence lifetime of each broom location in the human lens can be recorded, and thus the two-dimensional representation of the fluorophore as a cloth can be evaluated based on the fluorescence decay rate and intensity. Based on the fluorescence lifetime The decay-constructed color-coded image can be superimposed on the test image for clinical interpretation. It can be classified in the histogram by ^ all the photons corresponding to one pixel, and then: the private number sorrow function is fitted to the square root Lifetime information to complete the calculation of the fluorescent image. Then repeat this procedure for each pixel in the image. Soft = different method can use the tail fitting and numerical reconvolution to fit the data with the multi-index = variable function. The fitting procedure depends on the quality of the starting parameters used for the fitting. Therefore, the frequency count of the specific decay rate as an indication of the specific light-receiving group on the area scan can be directly determined by the image square. The aforementioned algorithm can be obtained by the electric moon: And can involve the presentation of data on a two-dimensional display such as a computer monitor. _ In one specific embodiment of the invention, the flatness associated with lifetime decay: intensity can be used as an aggregation of amyloid-like proteins. Measured, that is, the parameter can be used as a measure of aggregation by averaging the fluorescence intensity associated with the parameter pair and the decay of the sinus, for example, based on the parameter change, the progress of the disease in the body can be monitored by the computer. Or other specific hardware measurements. * The fluorescence histogram of the fluorescent starch-like complex obtained from the device of the specific month of this month: Compound (No. 11 compound) is shown in Figure 6. Single-counter number fitting is obtained 2 Lifespan decay rate of nanoseconds. Figure 7 shows the fluorescence lifetime image of the ut of the ut.: and its corresponding intensity image obtained according to the embodiment of the present invention. The image is 100 100 pixels obtained in 0.5 seconds and represents 5 〇.扫描 5〇_ scan area. One embodiment of the present invention uses fluorescence time domain technology to detect and analyze fluorophore m-intensity and lifetime metric based on lifetime characteristics, and obtain additional size information to identify fluorescent markers i In the test tube, the research proves that the specific example of the present invention is based on the ability of life decay characteristics to distinguish camp light fluorophores. In addition, the pharmacokinetics of rabbit eyes discussed in Experiment 2 A detectable fluorescent signal on the lens nucleus of the fluorescent starch-binding compound (No. n) is displayed. More importantly, the signal detected in the lens of the rabbit eye is easily recognized and belongs to the class (4) combined fit. The skilled artisan will understand that any of the methods presented herein (and their individual steps and combinations of several subsequent steps of such methods) relate in particular to the technical steps of collecting and processing the relevant data as appropriate, thereby obtaining data and normal control values. Comparison and/or discovery of any significant deviations during the comparison; prior to, subsequent to, and in preparation for subsequent separate diagnostic steps, ie, in the event of causing the obtained value and the target-like condition, such as The deviation between the normal control values (actual diagnosis) of Z-Mermoney's disease (in actual diagnosis), especially those that are performed before the subsequent separate diagnostic steps, independent of their 37 201215876, and when they are prepared for subsequent separate diagnostic steps ( Including individual steps and combinations of several subsequent steps of such methods) as individual specifics of the invention example. According to one embodiment of the present invention, an existing supplier can supply a plurality of sub-components of the system. For example, the excitation source can be PiC0Quant of Berlin,

Picosecond Pulsed Laser, LDH series sold in Germany; Picosecond Diode Laser, BDL series sold by Becker & Hickl of Berlin, Germany; or Picosecond Light Pulser, PLP series sold by Hamamatsu Photonics of Hamamatsu, Japan. Use Pic〇Quant,

TCSPC module sold by Berlin, Germany; Becker & Hickl, Berlin,

TCSPC module sold in Germany, SPC series; or Hamamatsu

Photonics, Hamamatsu, Japan For sale Synchronous Delay

Generator, Cl 0647 performs data acquisition. Photon counting detector can be

PicoQuant, Berlin, Germany Detector Unit for sale, PMA series, Becker & Hickl, Berlin, Germany Detector Unit for sale, ID-100 series; or Streakscope (cl〇627 series, Ham_tsu

Photonics, Hamamatsu, Japan for sale). It should be understood that other excitation sources, data acquisition modules, and photon counting detectors can be used. Portions of the above specific embodiments of the invention may be implemented using one or more computer systems. For example, specific examples can be implemented using hardware, software, or a combination thereof. When implemented in software, the software code can be executed on any suitable processor or set of processors provided in a single computer or distributed among multiple computers. In addition, it should be understood that the computer can be embodied in any of a variety of forms, such as a rack computer, a desktop computer, a laptop computer, a tablet computer, a circuit board computer or a system on a wafer. In addition, the computer can be embedded in a computer but has a suitable processing capability, including the number of individuals: (Daily Assistant, PDA), smart phone or any other suitable portable or fixed electronic device. Similarly, a computer can have one or more input and output devices. This: Especially useful for presenting user interfaces. Can be used to provide a user interface: Examples include a printer or display screen for visual presentation of output = a speaker or other sounding device for audio presentation output. Examples of input devices that can be used to enable the interface include keyboards and indicator devices such as mice, touch pads, screens, and digital input boards. The computer can receive input information by voice recognition or other voice types. 1 These computers can be used in any form by - or multiple networks: in the form of a local area network or a wide area network such as a corporate road. Such networks may be based on any suitable technology and may operate according to any ^= and may include wireless networks, wired networks, or fiber optic networks. The sigma scheme is similar, the guest insertion t, soil + j· __ | & method or processing described in this paper can be encoded as software executable on the processor, the processor uses a variety of operations ^ or platform Either. In addition, the software can be written using any of a variety of stylized s and / or stylized or scripted tools, and can be coded as $ ^ on a virtual machine in box (4). w 5 code or intermediate in this respect, at least a part of the present invention 5, 丨, . 叽 可 can be embodied as computer readable 经 coded by the program code; Read media) (eg, electricity 39 201215876 •: remember =, one or more floppy disks, compact optical discs, flash memory, field programmable gate arrays or other circuit configurations in a tomb detector, or Other tangible computer storage media), when coughing is performed on more than 22 computers or other processors, implement the implementation: = at least part of the specific instance. Computer readable media == The program stored thereon can be loaded to - or a plurality of different computers - & J sι to implement the various aspects of the invention described above. - In this respect, it should be understood that at least a part of the above-described specific examples includes at least one computer program (for example, a plurality of computer-readable computers), and the execution of the specific code (--Function. As used herein, the term "computer-readable medium") covers only computer-readable media that can be considered as a machine or manufactured: "items". Computer-readable media can be = coded or stored A tangible medium for computer readable information, a storage medium that can encode or store information, and/or a computer readable body that can be encoded or stored::::!media (__transit°ry medium). Examples include computer memory (such as ROM, Ram, flash memory or other types of computer memory), disk or tape, light, or other types of computer readable media that can be considered as machines or articles of manufacture. "Pr0gram" or "Software ("""""""""""""""""""""""""""" Or computer The instruction set can be executed. In addition, it should be understood that according to one aspect of the specific example, the executable or the evening computer program does not need to be resident in a single computer or processor, that is, 40 201215876 can implement the method of the present invention, and can be in the form of a module. Assignment to many different computers or processing of crying I, I k seems to be a continuation of many aspects of the invention. Computer-executable instructions can be executed by - or multiple computers or other devices. ^, such as a program module. In general, a program module includes a routine, a program, an object, a piece of data structure, etc. that implement a specific abstract data type. Typically, the program module can function as many Intra-in vitro study of a Neuropterix fluorescent ligand having a β-class powder (1-42): According to the invention, the _JL transition / / is taken in a specific example The test tube was incorporated into the test tube of the No. 11 compound Αβ peptide. The sputum study was carried out. Using the specific example of the present invention, the fluorescein lifetime measurement of the compound No. 11 of the Αβ peptide was used. For display according to This is a combination of the 广β-peptide of the Αβ peptide, the compound No. 11 and the light-life life histogram c camp light lifetime image. The map shows the decay rate....: Taizhong. By fitting Photon detection area with low lifetime and low level of life: II. The specific example of the invention can be more effective than the fluorophore with a difference of 0.85 nanoseconds. The experiment is described as follows. The purpose of this in-vitro study is the optical characteristics of the compound, and the characterization of: compound compound powder combination. In particular, the research objectives are: 1) the glory variability of the table (four); and 2) inspection, The ability of Compound No. 1 to be the compound of No. 11 of the dying. No, ... is combined with a quinone type starch (Αβ) peptide device: 41 201215876 Price (four) ring SAPPHIRE 11 device (NeUroptix Corporation,

Ac(10), MA'U.S.A.) is a special device for human clinical studies that is suitable for measuring this experiment in vitro. It employs a confocal scanning mechanism combined with a light life life spectrum that allows for the distinction of glory groups. The device allows the beam to be scanned to a specific location in the anterior segment of the eye (such as on the nucleus) and 2) to identify the fluorescent fluorophore based on fluorescence lifetime measurements. Method - Aggregated Aβ peptide preparation: The aggregated AP peptide was damaged by dissolving Aβ (1·42) in pBSpH7 4 until the final concentration was just μΜ. This solution was magnetically mixed for 3 days at room temperature under the condition of _. (10) _ ^ (1_42) in the _ _ reserve ♦ liquid and so on and frozen at -80 C for up to 4 weeks, and its characteristics without any improvement, 150 pL of aggregated Αβ (1-42) was added to 2.8 5 In mL No. 11 compound, 5 μΜΑρ(1_42) and 4_帛11? tiger compound were dissolved in a final concentration of 1 (4) and transferred to a 5 mL vial and subjected to hunger. EXPERIMENTAL AND RESULTS Samples of Compound No. 11 were placed in front of the Neuropxx §apphire =+ device. Once the scan position in the sample is determined, the grating is known to obtain a fluorescence lifetime measurement. Fig. 8A shows the image of the scanning range 100 μιη X 1〇〇μηι which is known in the acquisition time of 1 second (2〇〇x 2〇shadowing denier/m ^ ; the shirt image is not demonic decay. It constitutes the background of most images. The shading indicates a lifetime of 4 seconds, which corresponds to the lifetime of the fluorescent starch-binding compound, and the point detected in the image is a lifetime decay of 2.25 nanoseconds: aggregated Αβ peptide. The graph of Figure 8B shows No. 化合物 compound and human 42 201215876 Fluorescence decay rate calculated from compound No. n of aggregated Αβ peptide Conclusion In-tube fluorescence lifetime measurement of Compound No. 11 with aggregated ruthenium peptide was performed with a NeUroptix sAPPHIRE device. Based on the fluorescence lifetime decay rate, two peptides can be resolved with or without binding to the U-th compound. The results show that only the detection level of hundreds of photons can be used to distinguish the SAPPHIRE II with a lifetime of fluoroparticles with a difference of 85 nanoseconds. The superior efficacy of the device. Experiment 2: ocular pharmacokinetic study in a Dutch-white rabbit (DUtch-Belted Rabbit): According to one embodiment of the present invention, one of the present invention is used The device performs an in vivo pharmacokinetic study of Compound No. 11 in rabbit eyes. Figure 9A is a plot of photon frequency counts for two rabbits administered with starch-like binding compounds and for specific decay rates measured in rabbits and rabbits. According to an embodiment of the present invention, the photon frequency of the ratio of the decay rate of the fluorescence-like compound, the compound of the '° mouth compound (the compound No. 11) is calculated from the glory histogram (Fig. 9B), This demonstrates the ability of a starch-like binding compound to penetrate the cornea and be detected by a device in the lens of a rabbit's eye. The experiment is described below. Introduction The dose-response eye for topical administration of a fluorescent starch-binding compound in Helan•white-shoulder rabbits Pharmacokinetic study. Rabbits were tested daily for 4 days using Compound No. 11 starch-binding compound in the form of an ointment. Two animals were in the right eye in the form of ointment No. 11 compound 4 (0.5%) 衿荜. ; one-ft -^L· 4-/ . 〇', initially untreated and used as a control animal. The animal is administered at a specific time point for 4 τ

Fluorescence intensity and lifetime measurements were tested at the beginning and end of each day using the SAPPHIRE; II 43 201215876 system. The results showed that: 1) The detectable fluorescent signal was achieved at a concentration of 5 mg/g of the 11th compound in the form of an ointment after repeated topical administration. 2) Fluorescence measurement performed at the beginning and end of each day for 4 days showed an increase in fluorescence of Compound No. 11 in the lens nucleus of rabbit eyes. Method: Eye measurements were performed at time intervals and dose concentrations as described in the table below. Group treated animal number toxicant dose host eye measurement 1 5 mg / g (ointment) 2 4 days of ointment administered at 830, 1130 and 1430. SAPPHIRE II test control at the beginning and end of each day - 1 starts every day And at the end of the SAPPHIRE II test table 1: Test animal groups, doses administered and measurement time studies were staggered within 4 days and performed in a single location and on a single instrument. The test was conducted in a dimly lit room dedicated to the study. All animals were dosed with topical ocular administration as Compound No. 11 and tested in the right eye. Animals were anesthetized and manually held on the platform at the front of the Neuropterix SAPPHIRE II unit. The total positioning was performed by the animal processor and the position was fine-tuned by the Neuroptix SAPPHIRE II operator. Once aligned, the SAPPHIRE II operator initiates the measurement process. Animals were baseline measured prior to dosing and then at the beginning and end of each day (Table 1). Experimental Design and Dosage: Performs fluorescence lifetime and intensity in the eye at the beginning and end of each day. 44 201215876=ΓAxial scan (z scan) rabbit eye to obtain intraocular information and t=:xy scan) in the eye The specific area (in this case, the celestial body nucleus) performs a life decay variable test. ...:Identify the location of interest', that is, the start time related single photon count (TCSPS) when performing the scan. » Then the corpse is always on the top eight. You are the 仵 光 奇 奇 , , , The decay of the poisonous lun ji, the singer can be a histogram. The calculated decay time is the color coded in the life image. Each measurement was performed three times. From the xy scan, the decay rate of the compound No. 11 of the X-ray is calculated as the ratio of the decay rate: the sub-frequency count and the average of the three-dimensional measurement. Fluorescent dyes are introduced into the light-human rights test every day and show no significant drift during the entire period. *Monthly t* The device is specially designed for human use, and the platform is modified to hold rabbits. Administration was performed by topical administration to the right eye of each animal by a Toxikon staff. Animals were anesthetized with subcutaneous injections of Dexdomitor (〇5 mg/kg) and Ketamine (5 mg/kg). Then, an ointment tape of about 1/2 hour was applied to the right lower eye of each animal of the test group three times a day for 4 days 0... and, for the control group, one animal was in the same manner as the animals in the test group. Treatment, but no ointment or solution was administered to the eyes. Summary of results: The mother's 4 (in the morning) and the end (evening) performed the camping light intensity and life measurement of the powder-like compound in the eye. Figure i〇a and Figure _ are curves showing the photon frequency count of the specific decay rate of Compound No. 11 201215876, in which the baseline values were measured in the morning during the 4-day study period in 5 rabbit lens nucleus Measured at the end of each day after dosing. Figure 丨〇A is a graph of morning measurements, and Figure i 〇B is a graph of night measurements, both measurements were performed on rabbits administered with Compound No. 11 ophthalmic ointment. The measurement of the two rabbits (丨〇〇2 and 丨〇〇3) administered with the ointment ophthalmic ointment showed a significant increase in the fluorescence signal in the nucleus of the eye. Fig. A and Fig. B show that cumulative fluorescence signals were measured along the 4-day study period after three administrations at three hour intervals per day. Figures 11A and 11B show two fluorescence lifetime images acquired using animal 1〇〇3 after the baseline and fourth day studies. Baseline measurements reveal a black image indicating the absence of the 11th compound in the lens of the eye. After 4 days, the xy scan reveals: the life image of the decay rate of 2 nanoseconds (in gray), the characteristic of the fluorescent life ρ. The difference between the signals collected between the two rabbits may be attributed to the technician's Drug changes and animal flashing. Conclusion The quinone compound ophthalmic ointment of 払X 5 mg/g concentration was achieved. Repeated topical administration of Compound No. 11 tends to accumulate in the nucleus after a few hours of application and tends to stay for at least 12 hours. The teachings of the present application and the teachings of the present invention are not limited by the specific examples of the present invention. mouth. BRIEF DESCRIPTION OF THE DRAWINGS 46 201215876 FIG. 1 is a schematic view of an optical system according to an embodiment of the present invention. 2A is a graph of camping light intensity versus displacement measured during the calculation of the lens interface in the z-scanning eye, and the graph is a graph of FIG. 2A according to a specific example of the present invention. A graph of the first derivative. 3A and 3B are graphs illustrating the measurement of the fluorescence decay time according to an embodiment of the present invention. 4 is a schematic diagram illustrating the use of a time-correlated single photon count in accordance with an embodiment of the present invention. Fig. 5 shows the structure of the compound No. 11, which can be used as a fluorescent type starch-binding compound of the present invention. Fig. 6 is a fluorescence histogram of a fluorescent ruthenium-binding compound (No. 11 compound) obtained by a device of one embodiment of the present invention. Figure 7 is a diagram obtained in accordance with an embodiment of the present invention! A picture of the fluorescence lifetime image of the nickname compound and its corresponding intensity image. Fig. 8A is a fluorescence lifetime image showing a starch-binding compound such as a specific example of the present invention and a starch-binding compound bound to an aggregate peptide. Fig. 8B is a view showing a starch-binding compound such as the fluorescent ghetto image of Fig. 8A and a lake conjugated to the aggregate peptide according to an embodiment of the present invention. A plot of the corresponding fluorescence lifetime histogram of the compound. Fig. 9A is a graph showing the photon frequency of the specific decay rate of the fluorescent starch-based compound No. 11 measured in rabbits in an in vivo study according to an embodiment of the present invention. 47 201215876 Figure 9B is a fluorescent histogram according to the present invention. An experimental value corresponding to the specific example of the study of FIG. 9A and a compound of No. 1 after administration) FIG. 及A and FIG. 10B are shown in the present invention — _ 'About the morning during the rabbit study period A graph of the specific photon frequency of the fluorescence-based starch-binding compound measured at the end of each day. Figure UA and Figure 1 are at baseline and in animal studies. 1B is one of the present inventions.砰 丨 丨 — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — — Emission spectrum excited at 470 nm [Main component symbol description] 101: Array lens 102: sag photodiode detector (APD) 1 〇 3 : sample (eye) 104 : translation stage 1 〇 5 : current Mirror 402: sag photodiode (APD) 403: sample 406: pulse source 407: synchronous module (SYNC) 408: constant ratio discriminator (CFD) 4〇9: time-amplitude converter (TAC) 410: Multichannel Analyzer (MCA) 48

Claims (1)

201215876 VII. Scope of Application: · 1 · A method for detecting a starch in a mammalian eye, the device comprising: a light source configured to emit light to the eye and having a wavelength of light, *: The light of at least one of the combinations, each of which is suitable for: when the type of starch-binding compound is bound to the starch-like egg, the egg-like person A can kill the powder: two and: the cockroach produces the light A starch-binding compound has been introduced into the eye and specifically bound to the type of lake and π曰, which is indicative of an amyloid-like condition, configured to receive, including, illuminating the eye by illuminating the eye and determining at least Incorporating the fluorescence decay rate of the glory that is not associated with the genus of the genus of the genus, the determination allows the decay rate to be recognized in the genus of the genus The presence of the eye in the eye. 2. The device of the patent scope, wherein the optical unit (four) states to determine the decay rate of at least one of the following: molecules, compounds; Congo red or Congo & a Mp; / powdered humanized 厶 this., and Congo red Ho bio-starch ° 〇 ''Chrysamine' type starch-binding character. Gold main (four) powder binding compound; Golden amine G or Golden Jg Yan a compound; thioflavin tau or thioflavin derivative class II::: substance TM 5 or dynamin s-powder... 3. device as claimed in item i or item 2 Wherein the optical early indica is determined by the fluorescence intensity of the fluorescent light produced by at least the starch-binding compound 49 201215876 bound to the amyloid. 4. The application of the third paragraph of the patent application is ^ ^ ^ ^ Aligning, wherein the optical unit is configured to determine the type of the starch-binding compound conjugated to the amyloid based on the intensity and the decay rate of the time, 5. Or a device of the 2nd M «-z.., wherein the optical device is configured to determine a specific number of photons of the eye. * A device having a level of sorrow rate, wherein the light source is 6. For example, the first or the first part of the patent application scope includes a pulsed laser. The device of item 6, wherein the pulsed laser passes through the group 'I to emit light having a repetition frequency between about 1 MHz and about 24 () mHz. 8. And wherein the pulsed laser is configured to emit light having a pulse width between about 40 picoseconds and about 4 picoseconds wide. ▲, 9. The device of claim 6 wherein the pulse The laser is configured to emit a repetition rate of about 4 〇 MHz and a pulse width of about 2 〇〇 picoseconds. 10. The apparatus of claim 2 or 2, further package 3 configured An optical scanning unit that scans light from the source at the location of the eye. 11. The device of claim 10, wherein the optical scanning unit L3 is mounted on the translation stage and the scanner including the galvanometer mirror. 12. The device of claim 10, wherein the optical scanning unit 70 is arranged in red to sample at least one of the 50 201215876 injection areas of the eye using illumination of the light source, the sampling comprising illuminating a point in at least one region, At least one of a plane or volume and detecting fluorescent emissions. 13. The device of claim 1, wherein the optical scan is read by a single reading to sample different locations on more than one area of the eye. 14. The device of claim 1G, wherein the optical sweeping readout arrangement performs a planar scan of the eye using the light source in a continuous plane extending along a depth axis extending deep into the eye. 15. If the device of claim ii or item 2 is applied, the further step comprises a photodetector unit for detecting fluorescence emitted from the eye. 16. The device of claim 15 wherein the photodetector unit comprises at least one of a photodiode, a photomultiplier, a charge transfer device, and an enhanced charge coupling device. A device of item 15, wherein the photodetector unit 17. The refractory photodetector is included in the scope of the patent application. 18. The device of claim 15 further comprising a time-correlated single photon counting module that receives an electrical signal from the photodetector unit indicative of a photon count from the fluorescent light of the eye. 19. The device of claim 18, further comprising at least one processor module configured to determine a time decay rate of the fluorescence based on a photon count distribution that varies over time channel elements. 20. If you apply for a patent range! Item or item 2, wherein the optical unit is configured to distinguish between spontaneous phosphorescence, other non-specific particles, and unbound powders of the starch-binding compound and the ocular tissue from the starch-like egg Spontaneous fluorescence of the bound compound. 51 201215876 wherein at least one of the light is of the type 21; as in the device of claim 1 or 2, the unit is configured to identify the presence or amount of one or more of the following: a starch-like binding compound; a starch-binding compound bound to the amyloid; and an amyloid. 22. The device of claim 1 or claim 2, wherein the powder protein comprises aggregates. 23. The device of claim 1 or 2, wherein the amyloid comprises pre-amyloid aggregates. 24. If the scope of the patent is declared 帛! The device of item 2, wherein the amyloid protein comprises a beta starch. 25. The device of claim 2, wherein the amyloidogenic condition comprises Alzheimer's disease. 26. The device of claim 2, wherein the optical unit is configured to discriminate at least the presence of the amyloid based on the detected fluorescence. A device according to claim 1 or 2, wherein the optical unit is configured to determine the rate of delivery of the starch-binding compound to the eye based on the detected fluorescence. 2. The device of claim 1 or 2, wherein the optical unit is configured to determine a spatial distribution of the starch-binding compound delivered to the eye based on the detected fluorescence. 2 9. The device of claim 1 or 2, wherein the e-lighting unit is configured to determine a concentration gradient of the starch-binding compound at the corneal interface of the eye based on the detected fluorescence. 52 201215876 30. The device of claim 1, wherein the optical unit is configured to determine a spatial distribution of a compound of the diarrhea powder in the aqueous humor of the eye based on the detected fluorescence. And at least one of the time distributions of such starch-binding characters. σ 31_ as set forth in claim 帛i or item 2, wherein the optical unit is configured to determine an eye interface of the eye based on a fluorescent signal emitted by the tissue of the eye, νΛ g | position. 32. The device of claim 1 or 2, wherein the optical unit is configured to determine a position of the eye of the eye based on at least one of: (i) a distance from an anatomical structure of the eye And (Η) detection of changes in the intensity of mass production. 33. The device of claim 2, wherein the coughing optical unit is configured to be based on the anatomical structure or substructure of the eye, a small-part of the natural f-light excitation (four) anatomical structure or The substructure is up to two dimensions. 34. The method of claim 33, wherein the at least one dimension comprises at least one of: a thickness of the structure or substructure, a shape of the structure or substructure, and - or more : The distance between a structure or a substructure. 35. The device of claim 1 or 2, wherein the learning unit is configured to scan within the eye to determine the excited natural fluorescent light&apos; and thereby determine at least one region of interest in the eye; And using the illumination of the light source to sample at least one of the regions of interest in the eye / sampling I 3 to perform at least one complete region of the at least one region 53 201215876 wide measurement or use the flower, gg such as nine The original ..., the shot samples at least one of the _ positions in the at least one region - the sampling of the different locations includes at least one of a point, a plane or a volume in the same region; / to a sampling system for eliminating 丨# $^丨, &amp; 巧 了 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 由 。 。 。 。 。 。 。 。 。 。 。 。 。 。 'Claim 36. The device of claim 1 or 2, wherein the learning unit is configured to determine an axial sweep entering the eye along the depth: the natural luminescence of the excitation at each point, and thereby determining At least - the position of interest; and ^ wherein the optical unit is configured to determine a set of = surface scans in at least a continuous plane perpendicular to the direction of the axial scan of the eye using the source The fluorescence intensity of the fluorescent light and the time decay rate of the fluorescent light produced by the powder-binding compound of the amyloid of the amyloid at each point. 37. The device of claim i or item 2, wherein the device is configured to enable immediate search for the amyloid indicative of the halloid-like condition in the eye. 38. The device of claim 2, wherein the light source is configured to emit a peak region having a fluorescence excitation spectrum for the starch-binding compound bound to the amyloid in the eye. Light of the appropriate wavelength, and wherein the optical unit is configured to detect light having an appropriate wavelength for the peak region of the fluorescent emission spectrum of the starch-binding compound bound to the amyloid in the eye. 39. If you apply for a patent scope! Or the device of item 2, wherein the class of the 2012 20128876 starch binding compound is the ninth compound. 4. The device of claim 38, wherein the peak of the excitation spectrum is about 470 nm, the light source is configured to emit light at a peak of the excitation spectrum plus or minus about 20 nm, and wherein the emission The peak of the spectrum is about 58 〇 nm, and the optical unit is configured to detect light in the peak of the emission spectrum plus or minus about 2 〇 nm. 41. The device of claim 2, wherein the amyloid is an indication of an amyloidogenic condition. &lt; 42. A method for detecting an amyloid protein in a mammalian eye, the method comprising: illuminating the eye with a light source having at least one of a wavelength characteristic, a polarization characteristic, or a combination thereof, the characteristics being appropriate In the case of at least one class of powdered human compound bound to the amyloid in the class of powder-binding compounds: fluorescing 'this class of powder-binding compounds have been introduced into the eye and Terry &amp; An amyloid of the amyloid-like disorder; 'σ receiving light, including fluorescence generated by illuminating the eye; and measuring the firefly produced by at least the starch body of the starch conjugated to the _y# When the fluorescence of light is n 妄 玄 、, '〇口化 S 予 哀 率 , , , , , , , , , 该 该 该 该 该 该 该 该 该 该 该 该 该 该 , , 结合 结合 结合 结合 结合 结合 结合 结合 结合 结合 结合 结合 结合 结合 结合 结合 结合 结合The presence of the binding compound in the eye. early.. 〇化 43. As claimed in the Scope of the Patent No. 42 义 万 其 其 其 其 其 其 其 其 其 其 其 其 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 泪 至少 至少 至少 至少 至少55 201215876 The intensity and the decay rate of the time determine the amount of the S-type powder binding compound bound to the starch protein. The method of any one of claims 42 to 44, wherein illuminating the eye with the light source comprises illuminating the eye with a pulsed laser. 46. The method of any one of claims 42 to 44, wherein the method comprises irradiating the eye with light having a repetition rate between about .MHz and about 240 MHz. 47. The method of any one of claims 42 to 44, wherein the inclusion comprises exposing the eye to a light having a pulse width of from about 40 picoseconds to about 400 picoseconds. 48. The method of any one of claims 42 to 44 which comprises illuminating the eye with light having a repetition rate of about 4 〇 MHz and a pulse width of about 200 picoseconds wide. 49. The method of any one of claims 42 to 44, further comprising: increasing the position of the eye interface of the eye based on a fluorescent signal caused by natural fluorescence emitted by the tissue of the blinking eye . The method of any one of claims 42 to 44, further comprising: using the light source as one of the eyes, one of the regions of interest, the sampling comprising illuminating the at least one region At least one of a point, a plane, or a volume in the middle and detecting the fluorescent emissions. 5 1. The method of claim 5, wherein the sampling comprises sampling or different locations on more than one area of the eye. The method of any one of the preceding claims, further comprising: performing the eye using the light source in a continuous plane extending to a vertical axis in the eye depth-by-depth Plane scanning. ,,, 53. If the scope of the application for patents is 42 to 44, the progress of any one of the members includes: based on at least the following: the position on the nucleus of the eye: (丨) and the anatomy of the eye Structure and (variation of detection. Also 5: For example: please refer to any of the 42nd to 44th items of the patent scope: the powder-protein combination of this type of powder protein Eye group 2; the spontaneous fluorescence of the compound powder combination compound of the temple powder combination compound, the non-specific particles and the unfinished method, the patent application scope 42 to 44: the party of the item: Included in the presence of one or more of the following: _ ΉΠ 丨 子 子 子 子 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊 颊Class method = please patent scope ... to the 44th item - the party x class powder protein contains aggregates. Law: such as: please patent range 42 to 44, the term - the party 58. ; 1:: The protein contains the former class of powdered protein aggregates. The French patent range 42 to 44 ε ^ where the _ powder protein contains &quot;please powder. 财任-方之方Μ. If you apply for patent scopes 42 to 44, _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ _ The method of any one of claims 42 to 3, wherein the powder binding compound comprises a molecular rotor. 61. If the patent application range is 42 to 44 The method of any one of the present invention, wherein the starch-binding compound comprises at least one of the following: Congo red or Congo red derivative starch-binding compound; a golden amine starch-binding compound; a golden amine derivative-based starch-binding compound; Or a gold amine G derivative I powder binding compound; thioflavin tau or thioflavin T derivative class powder binding compound; and thioflavin s or thioflavin s derivative; Compound 62. If the patent application scope is 帛42 to 44, the 3 is based on the detected fluorescence to discriminate at least the presence of the powder of the genus. 63. Item to item 44 A method of further comprising the method of detecting the rate of transmission of the starch-bound character of the genus A. 疋 疋 。 。 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如 如The method further comprises determining the average number of photons at the eye rate. The specific combination of the items of the 44 items of the damage to the eye, such as any one of the 44 items, is determined to be a combination of the starches of the type of the starch. For example, in the scope of claim 42 to the method, it further comprises a spatial distribution based on the detected fluorescence-measuring powder-binding compound. 66. If the patent scope is in the 42nd to the third, the method further comprises a detection-based fluorescence measurement. 58 201215876 Concentration gradient at the interface of the angle of the eye. 67. The method of any one of claims 42 to 44, further comprising the spatial distribution of the pulverized binding compound in the aqueous humor of the eye based on the detected fluorescence and the like At least one of the time distribution of the starch-integrated character. </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; </ RTI> <RTIgt; Structure or sub-social structure is less than one size. And 69. The method of claim 68, wherein the method is as follows: at least one of the following: measuring the thickness of the structure or substructure, determining the weight of the structure or substructure, The shape of the structure and the distance between one or more structures or substructures of the eye. 7. In the case of any one of the 42nd to 44th patent applications, the further step of detecting the fire generated by the eye using a photodetector device is as follows: The method, wherein the photodetector comprises at least one of a photodiode, a photomultiplier, a charge coupled device, and a fused device. 72. The method of claim 1, wherein the photodetector comprises a fast burst photodiode detector. Method: 3· For example, please refer to the party of any of items 42 to 44 of the patent range ^ ^Pack 3 to perform the time-dependent single-sub-count of the fluorescence produced by the eye. 59. The method of claim 73, wherein performing the time phase D° photon counting comprises measuring the time decay of the fluorescent light in a pulsed form and based on a photon count distribution of a single channel change over time channel rate. The method of any one of claims 42 to 44, comprising: determining the scan in the eye to determine the excited natural fluorescent light and from at least one of the regions of interest in the eye; Detecting at least one of the regions of the eye using the illumination of the source of interest by the illumination of the source of interest. The sampling comprises illuminating at least one of the points in the at least one region; the sampling is for determining at least s-mysterin The light intensity of the starch and the time decay rate of the fluorescent light. 76. The method of claim 42, wherein the method comprises: the fluorescent component of the fluorescent compound generated by the binding compound in the at least one sampling region to the depth of the axis in any one of the 44th The excited girl at each point shakes out η μ ~ eight hot 且 light and thus measures the position of interest; and one less uses the light to align the plane of the eye in a continuous plane perpendicular to the axial scan of the ancient a M 13 Broadcasting 70 to determine the fluorescence intensity of the fluorescence of the starch-binding compound bound to at least the powdered protein protein at the plane scanning points and the time of the fluorescein produced by the fluorescent t _ π Decay rate. Act 77. If the scope of the patent application is such that the method enables any of the 42-to-44th items in the eye to be searched for in the eye, the amyloid of the class of the 2012 20128876 powder-like condition. 78. The method of claim 228, wherein the method further comprises the step of having a fluorescence excitation spectrum for the starch-binding compound bound to the amyloid in the eye. Light of a region of appropriate wavelength illuminates the eye; and 'field detection of the wavelength of the peak of the fluorescence emission spectrum of the starch-binding compound bound to the amyloid in the eye Light. 3 ^ ^ 79. The method of claim 78, wherein the starch-binding compound is the compound No. 11. 8. The method of claim 7, wherein the peak of the excitation spectrum is about 470 nm, and the illumination of the eye is at a peak of the excitation spectrum plus or about a wavelength within about 20 nm, and wherein the emission The peak of the spectrum is about 58 Å, and the light received by the eye is detected at the wavelength of the emission spectrum plus or minus the wavelength within about 2 〇 nm. 81. The method of any one of claims 42 to 44 wherein the amyloid is an indication of an amyloidogenic condition. The method of any one of claims 42 to 44, wherein the method allows at least two different fluorophores having a similar fluorescence spectrum to be distinguished based on the decay rate at the time, the similar fluorescence spectrum At least one of a significant overlap in the emission spectrum and the excitation spectrum is included. The method of any one of claims 42 to 44, further comprising the distribution of at least one of fluorescence intensity and lifetime decay of at least one fluorophore in two dimensions. 61 201215876 84. The method according to any one of claims 42 to 44, wherein the step further comprises determining the number of combined photons based on at least one of a camping light intensity and a lifetime decay of at least one fluorophore And the number of photons that are not combined. The method of claim 84, which further comprises the distribution of the fluorescence intensity and lifetime decay of the starch-binding compound which binds to the protein and the starch-binding compound which is not bound to the protein in two sizes. 86. The method of claim 85, further comprising synchronizing the performance of the two sizes with at least one of a scanner and a laser. 87. The method according to any one of claims 42 to 44, wherein the step comprises determining the parameter by averaging the fluorescence intensity associated with the decay of the specific lifetime over a particular region of the eye. The method of any one of claims 42 to 44, further comprising aligning the alignment source with the eye along a confocal path to determine a reference point in the eye. 89. A method for identifying an anatomical structure in a mammalian eye, the method comprising: illuminating the eye with a light source having at least one of wavelength characteristics, polarization characteristics, or a combination thereof The characteristics of the temple are each suitable for producing natural fluorescence in the anatomical structure of the far eye; and the intensity of the natural fluorescence produced by the illumination of the light source in the eye of the eye-eye is changed according to the natural The location where the change in fluorescence intensity is greatest identifies the anatomical structure. 〇·如申α Patent Section 89 of the method' wherein the anatomical structure 62 201215876 contains _### in front of the eye. Wherein the anatomy is the largest of the natural body fluids at least the body and the core 91. As claimed in claim 89 or the identification of the material structure comprising the 敎" „ position. 92. The method of claim 91, wherein the determining comprises determining an interfacial position of the lens capsule of the eye based on determining an increase in the intensity of the natural f light. 93. The method of claim 89 or 90, wherein identifying the anatomical structure comprises generating at least one of the following in the eye based on the light source: corneal thickness, corneal shape, eye shape The body shape, the thickness of the lens, and at least one of the thickness and shape of a substructure of the lens of the eye. 94. The method of claim _ 931, wherein the at least one substructure comprises the lens capsule, the cortex, and the nucleus of the eye: one less. 95. The method of claim 89, wherein the identifying the cross-sectional structure comprises determining an intraocular distance between at least two anatomical structures of the eye. 96. The method of claim 89, wherein the method further comprises detecting, by the light source, a powder protein indicative of the amyloid-like condition in the eye of the mammal. 97. The method of claim 96, wherein the method further comprises: illuminating the eye of the mammal with the light source, the light source further comprising at least one of a wavelength characteristic, a polarization characteristic, or a combination thereof, the characteristics Each suitable for producing a fluorescent in the starch-binding compound when at least one starch-like binding compound binds to the amyloid indicating the starch 63 201215876-like condition has been introduced into the eye and specifically binds Such an amyloid indicating the amyloidogenic condition; receiving light, including fluorescence generated by irradiation of the eye; and measuring fluorescence of at least the fluorescent light produced by the starch-binding compound bound to the amyloid Time decay rate, the assay allows discrimination of the presence of such a matrix-binding compound bound to the amyloid-like protein in the eye based at least on the decay rate. a~々成, the presence of such a starch-binding compound in combination with the amyloid of the amyloid containing the starch-binding compound of the amyloid and the spontaneous fluorescence of the ocular tissue, other non-specific particles and Unincorporated temples spontaneously fluorescent. 99. The method of claim 89 or item % of the patent application &gt; means: enabling the amyloid in the eye to be immediately searched for; . Silver Μ 1 100• If the application of the patent scope, step 89, comprises illuminating the eye with a fluorescent excitation spectrum for the powder-binding compound in the eye; and the method of item 90, which is further integrated into the powder protein The light of the appropriate wavelength of the peak region of the temple has a wavelength of light that is detected by the fluorescent emission light of the compound in the eye. A method of binding to a peak region of such a starch spectrum. The method of claim 100, wherein the starch compound is a compound No. 11. 102. The method of claim 1, wherein the peak of the excitation spectrum is about 470 nm, and the illumination of the eye is added or subtracted by about 2 〇 nm at the peak of the excitation spectrum. At the wavelength within, and wherein the peak of the emission spectrum is about 5 ribs, the light received by the eye is detected at a wavelength of about 2 加 at the peak of the emission spectrum. 1 0 3. A method for determining a luminescent group in which the protein in the eye woven fabric is temporarily depleted, and the method comprises: having a wavelength characteristic, a polarization characteristic, or a combination thereof, The source illuminates the ocular tissue, and each of the properties is adapted to generate fluorescence in the genus powder-binding compound when at least the phyllo-powder compound is bound to the protein f, and the starch-binding compound has been incorporated into β hai The ocular tissue is specifically and specifically bound to the protein; the light includes: the camp light generated by illuminating the eye; and the camp light that is determined by at least the fluorescence generated by the conjugated compound of the protein bound to the protein &amp; Time bombing rate, #,一^ This test allows for the identification of the presence of binding to the protein tissue based at least on that time.夂 and age-like powder combination compounds in the eye VIII, schema: (such as the next page) 65